Below are the standard names and descriptions of the major plates involved in OCTOPUS 3.0:
| Name | Pupose | Type |
|---|---|---|
| Culture Plate | Contains 110 µl 2xYT + antibiotic to where colonies are picked into. Colonies grown overnight will be glycerol stocked with 100 µl 50% glycerol the following day. | Flat-bottom culture plates 96 well VWR |
| Sample Plate | Contains 25 µl water to where 5 µl of colony sample is added. Will be boiled and is input for OCTOPUS. | Skirted PCR plate 96 well BIORAD |
| RCA Plate | Contains 2 µl of pre-stocked Fast RCA reaction to where eventually 2 µl of Sample Plate will be added. This is the EquiPhi29 reaction (miniaturized 5X, modified with "focused" RCA). | 384 well PCR plate ENDURA |
| Ready Reaction Plate | Contains the i7 indexes that uniquely id each well up to 96. Wells should NOT be intermixed. | ExpressPlex provided plate 96 well |
| Indexing Plate | Contains the i5 indexes that uniquely id each set of 96 wells. Wells theoretically can be intermixed but best not to. | ExpressPlex provided plate 96 well |
| Dilution Plate | Contains 15 µl IDTE to where 1 µl of the incubated RCA Plate will be added (1:16 global dilution). It is crucial that this is thoroughly mixed, as RCA product can be quite goopy from the amount of DNA amplification. | 384 well PCR plate ENDURA |
| Reaction Plate | Contains the 2 µl Ready Reaction + 1 µl Indexing Plate reagents to where 1 µl of the Dilution Plate will be added. This is the ExpressPlex reaction (miniaturized 4X) that will produce the NGS libraries. Will eventually be placed on the OT-2 for pooling and purification. | 384 well PCR plate ENDURA |
| Reagent Plate | Contains reagents involved in purifying the NGS libraries. This is placed on the Magnetic Module on the OT-2 where the Reaction Plate will be pooled. | 2ml V-bottom deep well plate NEST |
| DNA Plate | This plate is placed on the OT-2 where it will contain the intermediate pooling step. After purification on the Magnetic Module, the purified libraries will be transferred back to this plate. | Skirted PCR plate 96 well BIORAD |